December 19, 16
Flow cytometry is a technique that allows researchers to identify and quantify different cell types within a sample by measuring the presence of different markers used to label the cells. The Ragon Institute Imaging Core Facility provides access to a range of different instrumentation that allows researchers to perform these characterizations using a variety of technologies.
Standard flow cytometry uses fluorescently-labeled antibodies and probes to stain the sample, and light signals are measured as cells are passed through different colored lasers to determine the amount of signal in each channel or parameter. The Ragon Institute maintains 5 different analysis flow cytometers (BD Biosciences), equipped with 2 to 5 lasers and capable of measuring up to 18 colors.
Sorting flow cytometers enable the researcher to collect cells of interest from the sample after “interrogation”, allowing for further studies of specific populations. We have a 5 laser, 18 color sorter that can collect up to 4 different populations at a time, housed in a biosafety cabinet to allow safe processing of infectious samples, and a 3 laser, 12 color sorter in BL3 able to collect 4 different populations at a time.
A technology that was only recently commercialized uses mass spectrometry instead of fluorescence to identify labeling, replacing fluorochromes with rare earth metals. This allows simultaneous detection of over 30 parameters at a time. The CyTOF (DVS Sciences) has great promise to provide more information per cell than ever before.
Imaging Cytometry combines the high-data content of microscopy with the high throughput of flow cytometry, capturing images of each particle in multiple channels instead of just quantifying the signal as in standard flow cytometry. The Ragon Imaging Core has a 12-channel, 4 laser imaging cytometer with multiple magnification options (The Imagestream X Mark II from Amnis).
Label-free interactions can be detected using our Biacore 3000 through SPR (Surface Plasmon Resonance). This technology can determine binding kinetics and protein interactions without the need for additional dyes or labels.
Luminex technology allows users to measure the presence of multiple proteins or other molecules in a sample, and the Ragon Institute now has an instrument capable of measuring up to 500 different analytes in a single sample.
Bulk sorting of populations using magnetic columns to select for positive markers can be performed on our AutoMACS cell separator (Miltenyi).
Mr. Michael Waring
Associate Director, Imaging Core-Flow Cytometry
Ragon Institute of MGH, MIT, and Harvard
400 Technology Square
Cambridge, MA 02139
Phone: (857) 268-7069
Dr. Galit Alter, Ph.D.
Director, Imaging Core
Ragon Institute of MGH, MIT and Harvard
400 Technology Square
Cambridge, MA 02139
The Ragon Institute Imaging core has an online electronic management tool. Users are required to sign up and provide funding information for the payment of usage fees. Contact Michael Waring for access.
The Ragon Institute provides training in flow cytometry and instrument use, through individual training sessions and introductory seminars. Registration required. Informational seminars will also be hosted periodically, check the “News” for upcoming events.
- 5 Laser LSR Fortessa, “5L”, 18 colors: 350 (2), 405 (6), 488 (2), 561 (5), 640 (3), HTS
- 4 Laser LSR II, “4L”, 18 colors: 350 (4), 405 (6), 488 (5), 633(3).
- 4 Laser LSR II, “4R”, 18 colors: 350 (2), 405 (6), 488 (6), 640(4).
- 3 Laser LSR II, “3L”, 16 colors: 405 (8), 488 (5), 640(3), HTS
- 2 Laser FACSCalibur, 4 colors (488, 633)
- BD SORP FACSAria cell sorter in biosafety cabinet, 5 lasers, 18 colors, 4 sort populations, small particle detection (350, 405, 488, 561, 640)
- Imagestream X Mark II Imaging Cytometer
- DVS Sciences CyTOF mass cytometer,
- Biacore 3000 label free protein interaction
- AutoMACS Pro cell separator
- Bio-Plex 3D Suspension Array System (xMAP technology)
The Ragon Institute Imaging Core is a member of the MGH Research Cores:
and the Harvard Center for AIDS Research (CFAR) Cores:
Core Facilities and Shared Resource Labs are assessed by the impact they have on the research conducted using their equipment and expertise. One metric of this is how many publications are supported by them. If you publish work using a core facility, please be sure to acknowledge their contribution to your work, so that you can continue to enjoy the benefits of their existence!
If you include an acknowledgement of our core, please send us the article citation, and you can use some version of this statement:
“The authors would like to acknowledge the contributions of the Ragon Institute Imaging Core Facility, which is supported in part by the Harvard University Center for AIDS Research (CFAR), an NIH funded program (5 P30 AI060354-10).”
The official statement from the CFAR can be found HERE.